Use of harringtonine in the preparation of a drug for treating hiv infectious diseases

Abstract

The application of homoharringtonine in the preparation of drugs for treating HIV-infected diseases belongs to the field of biomedical technology. This invention provides the application of homoharringtonine in the preparation of drugs for treating HIV-infected diseases; and a pharmaceutical composition for treating HIV infection, said composition comprising homoharringtonine and pharmaceutically acceptable excipients. This invention demonstrates through cell model experiments that homoharringtonine has a significant effect on inhibiting HIV invasion of cells and inhibiting HIV replication within host cells. This drug not only expands the selection of drugs for HIV infection but also further broadens the application scope of homoharringtonine.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology. It relates to novel pharmaceutical uses of homoharringtonine, specifically its application in preventing HIV infection and inhibiting HIV replication in infected individuals. Background Technology

[0002] AIDS (Acquired Immunodeficiency Syndrome) is an acquired immunodeficiency disease caused by infection with the human immunodeficiency virus (HIV). Since its first report in the early 1980s, AIDS remains a global health problem. AIDS not only has a significant impact on the physical health of infected individuals but also on their mental well-being. Currently, there are six types of drugs available free of charge in China: efavirenz (EFV), zidovudine (AZT), tenofovir (TDF), nevirapine (NVP), kaletra (Lopinavir / Ritonaviir), and lamivudine (3TC). In addition, there are more expensive out-of-pocket medications. HIV-infected individuals require lifelong medication, which leads to drug resistance; therefore, the search for new drugs is crucial.

[0003] Harringtonine is a natural Cephalotaxus alkaloid that inhibits protein synthesis and possesses antiviral activity against Chikungunya virus (CHIKV). To date, there are no research reports on the use of harmonium as a drug for HIV / AIDS.

[0004] In view of this, the present invention is proposed. Summary of the Invention

[0005] In view of the deficiencies in the prior art, the purpose of this invention is to design and provide an effective new anti-HIV drug, and the use of homoharringtonine in controlling HIV infection.

[0006] To achieve the above objectives, the present invention adopts the following technical solution:

[0007] Application of homoharringtonine in the preparation of drugs for treating HIV-infected diseases.

[0008] The application in question refers to HIV infection, specifically AIDS.

[0009] In the aforementioned application, the concentration of homoharringtonine in the drug is 0.1-5 μM.

[0010] In the aforementioned application, the concentration of homoharringtonine in the drug is 0.625 μM.

[0011] A pharmaceutical composition for treating HIV infection, the pharmaceutical composition comprising homoharringtonine and pharmaceutically acceptable excipients.

[0012] The pharmaceutical composition for treating HIV infection, wherein the concentration of homoharringtonine in the pharmaceutical composition is 0.1-5 μM.

[0013] The aforementioned pharmaceutical composition for treating HIV infection, wherein the concentration of homoharringtonine in the pharmaceutical composition is 0.625 μM.

[0014] Specifically, this invention conducts in vitro cell experiments on the drug homoharringtonine using Rev-A3R5-GFP cells and pNL4-3 HIV pseudovirus as the HIV packaging virus. Rev-A3R5-GFP cells utilize the interaction between HIV Rev and RRE (Rev Response Software) to regulate tracer gene expression, thereby overcoming the defects of the LTR reporter gene. Rev is only present in HIV-positive cells; therefore, the high specificity of Rev can significantly reduce background signals and increase detection sensitivity. Thus, Rev-dependent tracer cells can specifically detect low levels of HIV replication.

[0015] This invention uses the HIV drug nevirapine (NVP) as a positive control and analyzes the anti-HIV efficacy of different concentrations of homoharringtonine using flow cytometry. The results show that homoharringtonine has a significant inhibitory effect on HIV infection. Furthermore, the homoharringtonine described herein can be used to prepare drugs for treating HIV infection, demonstrating promising development and application prospects.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] 1. This invention discloses for the first time a novel use of holoharbine in the fight against HIV infection.

[0018] 2. This invention is based on Rev-A3R5-GFP cells and a pNL4-3HIV pseudovirus drug screening system. The inhibitory effect of the drug of this invention on pNL4-3HIV pseudovirus infection was analyzed by flow cytometry. It was discovered after screening a large number of molecules in the drug molecule library. There is no possibility that other compounds have provided any technical inspiration. Furthermore, the research and development of homoharringtonine has shown that it has a significant therapeutic effect on HIV infection. Attached Figure Description

[0019] Figure 1 The structural formula of cephalotaxine base;

[0020] Figure 2Flow cytometry plots showing the inhibitory effects of the virus control group, nevirapine control group, and homoharringtonine group on Rev-A3R5-GFP cells and the pNL4-3 HIV pseudovirus drug screening system;

[0021] Figure 3 The concentration-effect curve of cephalotaxine against HIV. Detailed Implementation

[0022] For ease of understanding, the present invention will be described in detail below with reference to the accompanying drawings and embodiments. It should be particularly noted that the following drawings only illustrate certain embodiments of the present invention. Those skilled in the art can make appropriate modifications and changes to the present invention within the scope of this invention based on the description, and these modifications and changes are also included within the scope of this invention.

[0023] The harmringtonine involved in this invention was purchased from Taoshu Biotechnology Co., Ltd. Harringtonine's CAS number is 26833-85-2, and its molecular formula is C2. 28 H 37 NO9, structural formula as follows Figure 1 As shown.

[0024] Example 1: Homoharringtonine inhibits HIV invasion of cells

[0025] Rev cells were resuscitated and cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, 100 units / mL penicillin, and 100 μg / mL streptomycin at 37°C and 5% CO2. Cells were passaged every 24 hours. Drug action experiments were initiated when cells reached the third passage and showed good microscopic appearance (Rev cell density reached 80%). The third and fourth passages were optimal for these experiments.

[0026] Cells were collected by centrifugation at 200G for 5 minutes. Rev cells were seeded at a density of 20,000 / mL in 96-well plates, with 100 μL of cell culture per well. The experiment included a blank control, a negative control, and a positive control. After seeding, 1 μL of homoharringtonine (0.2 mg / mL) was added to the wells of the experimental groups, while the positive control group received nevirapine, a clinically proven positive control drug. The blank and negative control groups received no drugs. After drug administration, the cells were incubated at 37°C for 1 hour.

[0027] After incubation, the experimental group was supplemented with 1.22 μL of homoharringtonine, the positive control group was supplemented with 1.22 μL of nevirapine, and 100 μL of pNL4-3 HIV pseudovirus solution and 22 μL of infectin were added to the negative control, positive control and experimental group. The blank control group was supplemented with 100 μL of culture medium. The mixture was incubated at 37°C for 4 h.

[0028] For the final administration, add the corresponding drug to the corresponding well at a volume of 8 μL, and then replenish the volume of each well to 1 mL with culture medium. The drug and virus interact with the cells for 48 hours.

[0029] After co-incubation, the cell solution in the 96-well plate was transferred to a flow cytometer and centrifuged at 400G for 5 minutes.

[0030] After centrifugation, the supernatant was aspirated, and 0.25 μL of Sytox dye and 100 μL of PBS solution were added to each tube for staining for 10 minutes. After staining, 350 μL of PBS was added to each tube, and GFP expression was detected by flow cytometry.

[0031] The results are as follows Figure 2 As shown, the left graph (virus control group (no drug)) shows that without drug treatment, the pNL4-3 HIV pseudovirus can achieve a GFP positivity rate of 18.8%. The middle graph (nevirapine control group (10 uM)) reduces the GFP positivity rate to 0.069%, indicating that the positive control drug is effective. The right graph (homonasine group (0.625 uM)) reduces the GFP positivity rate to 0.080%, and its effect on inhibiting pNL4-3 HIV pseudovirus is close to that of the positive control drug, significantly inhibiting HIV. It is evident that the positive drug nevirapine (nevirapine control group) can significantly reduce the GFP+ cell percentage of pNL4-3 HIV pseudovirus (virus control group), while honokiol has a similar effect (honokiol group).

[0032] In this embodiment, homoharringtonine was diluted at different concentrations to screen for suitable drug concentrations. Figure 3 The horizontal axis represents the concentration of holoharponine (µM), and the vertical axis represents the inhibition effect (the smaller the number, the better the inhibition effect; 0 represents complete inhibition); the results are as follows. Figure 3 As shown, 0.1 μM has a weak inhibitory effect, while 0.625 μM can achieve a similar inhibitory effect (0.080%) to the positive control drug nevirapine.

Claims

1. Use of harringtonine in the preparation of a medicine for resisting HIV.

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